Natural killer T (NKT) cells are an evolutionarily conserved subset of T cells that are developmentally and functionally distinct from conventional T cells. The ability to quickly secrete large quantities of a variety of cytokines upon activation enables NKT cells to be potent regulators of diverse immune responses. The deficiencies in NKT cell number and function have been linked to the development of many diseases. However, a significant gap remains in our understanding of how the development and function of NKT cells are precisely regulated. MicroRNAs (miRNAs), a recently discovered class of evolutionarily conserved small non-coding RNAs, negatively regulate the expression of protein-coding genes and thereby control essential biological functions and contribute to the development of many diseases. We were the first to report that the deletion of Dicer (a key enzyme for miRNA biogenesis) during hematopoiesis results in a significantly reduced NKT cell number and impaired NKT cell maturation and function, without alternating conventional T cell development in the thymus, suggesting that miRNAs are required for NKT cells. Our long-term goal is to understand how miRNAs regulate NKT cell development and function. While more than 1000 experimentally reported miRNAs, very few specific miRNAs are linked to NKT cells so far. Our objective here is to define specific miRNAs and their targets that regulate NKT cell development and function. Using miRNA arrays, we recently identified dynamic expression of miRNAs, including miR-155, and miR-17-92 cluster, during NKT cell development and activation. These findings plus our recent other report lead to our central hypothesis that these dynamically expressed miRNAs serve as critical regulators controlling NKT cell development and function through fine-tuning of specific target genes. Here we will further test this hypothesis. We will investigate how dynamic and miR-155 and miR-17-92 expression regulates NKT cell development and function using specific miRNA mutant mice with the gain or loss of miRNA gene. The results from proposed studies may not only illuminate the new immunological and molecular mechanisms underlying NKT cell development, but may also facilitate the development of new and more efficient intervention strategies for autoimmune diseases, infection, and cancer based on the NKT cell therapy.